Journal: PLOS ONE

Article Title: Protein production from HEK293 cell line-derived stable pools with high protein quality and quantity to support discovery research

doi: 10.1371/journal.pone.0285971

Figure Lengend Snippet: Analytical SEC summary of proteins 1–11 expressed in different expression systems.

Article Snippet: HEK293S GnTI - adherent cells (cat. no. CRL-3022, ATCC) were adapted to suspension cultures internally and grown in FreeStyle 293 expression media (cat. no. 12338018, Thermo Fisher Scientific).

Techniques: Expressing

Journal: PLOS ONE

Article Title: Protein production from HEK293 cell line-derived stable pools with high protein quality and quantity to support discovery research

doi: 10.1371/journal.pone.0285971

Figure Lengend Snippet: (A) Workflow of HEK293S GnTI - and Expi293F GnTI - stable pool generation via the PB transposon system. (B) Expi293F GnTI - kill curves of un-transfected cells with different puromycin (upper panel) and hygromycin (lower panel) concentrations. The chosen antibiotic concentration for selection was the lowest concentration that killed 100% of un-transfected host cells but not transfected cells (red arrows). (C) Viability (left panel) and VCD (right panel) showed that HEK293S GnTI - and Expi293F GnTI - expressing proteins 5 or 6 recovered in about 14 days after antibiotic selection. Mock and Neg refer to empty vector transfected and un-transfected cells, respectively. (D) Production harvest cell viability (left panel) and VCD (right panel) for HEK293S GnTI - and Expi293F GnTI - stable pools expressing proteins 5–10.

Article Snippet: HEK293S GnTI - adherent cells (cat. no. CRL-3022, ATCC) were adapted to suspension cultures internally and grown in FreeStyle 293 expression media (cat. no. 12338018, Thermo Fisher Scientific).

Techniques: Transfection, Concentration Assay, Selection, Expressing, Plasmid Preparation

Journal: PLOS ONE

Article Title: Protein production from HEK293 cell line-derived stable pools with high protein quality and quantity to support discovery research

doi: 10.1371/journal.pone.0285971

Figure Lengend Snippet: (A) Expression titer and (B-C) purification yield for proteins 5 to 10 expressed in CHO-K1 stable pools (blue bars), HEK293S GnTI - stable pools (red bars) and Expi293F GnTI - stable pools (green bars).

Article Snippet: HEK293S GnTI - adherent cells (cat. no. CRL-3022, ATCC) were adapted to suspension cultures internally and grown in FreeStyle 293 expression media (cat. no. 12338018, Thermo Fisher Scientific).

Techniques: Expressing, Purification

Journal: PLOS ONE

Article Title: Protein production from HEK293 cell line-derived stable pools with high protein quality and quantity to support discovery research

doi: 10.1371/journal.pone.0285971

Figure Lengend Snippet: (A) C-terminally His6-tagged protein 5 expressed from CHO-K1, HEK293, HEK293S GnTI - and Expi293F GnTI - was analyzed by SDS-PAGE under non-reducing and reducing conditions and stained by Coomassie blue. CHO-K1, HEK293S GnTI - and Expi293F GnTI - expressed protein 5 were derived from stable expression pools. Protein 5 expressed from HEK293 was from a commercial source (R&D Systems). For each cell line, 0.5 μg (left) and 1 μg (right) of protein 5 was loaded. (B) As in (A), but with protein 6 with a C-terminal His6 tag expressed from CHO-K1, HEK293S GnTI - and Expi293F GnTI - stable pools. The gel was visualized by stain-free imaging. Protein 5 (C) and protein 6 (D) expressed from CHO-K1 stable pools (blue), HEK293S GnTI - stable pools (red) and Expi293F GnTI - stable pools (green) were analyzed by analytical SEC. Zoom-in analytical SEC traces of protein 6 expressed from different cell lines are also shown in (D). (E) As in (B), but with protein 7 and protein 8 with a C-terminal His6. (F-G) As in (D), but with (F) Protein 7 and (G) Protein 8 and their respective zoom-in analytical SEC traces.

Article Snippet: HEK293S GnTI - adherent cells (cat. no. CRL-3022, ATCC) were adapted to suspension cultures internally and grown in FreeStyle 293 expression media (cat. no. 12338018, Thermo Fisher Scientific).

Techniques: SDS Page, Staining, Derivative Assay, Expressing, Imaging

Journal: PLOS ONE

Article Title: Protein production from HEK293 cell line-derived stable pools with high protein quality and quantity to support discovery research

doi: 10.1371/journal.pone.0285971

Figure Lengend Snippet: (A) The disulfide-linked two-chain protein complexes with a C-terminal His6 tag fused to one chain (protein 9 and protein 10) were expressed from Expi293F GnTI - , HEK293S GnTI - and CHO-K1 and analyzed by SDS-PAGE under non-reducing and reducing conditions. The gel was visualized by stain-free imaging. Clipping was observed for the CHO-K1-produced protein. (B) As in (A) but with proteins 4, 9 and 10 expressed from CHO-K1 stable pools. The gel was visualized by stain-free imaging. * indicates clipped species of protein 4. (C) Protein 9 and (D) protein 10 expressed from CHO-K1 (blue), HEK293S GnTI - (red) and Expi293F GnTI - (green) was analyzed by analytical SEC. Zoom-in traces are also shown.

Article Snippet: HEK293S GnTI - adherent cells (cat. no. CRL-3022, ATCC) were adapted to suspension cultures internally and grown in FreeStyle 293 expression media (cat. no. 12338018, Thermo Fisher Scientific).

Techniques: SDS Page, Staining, Imaging, Produced